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Cloning and Sequence Analysis of a cDNA Encoding Ferric Leghemoglobin Reductase from Soybean Nodules

AutorJi, Lin; Becana Ausejo, Manuel ; Sarath, Gautam; Klucas, R. V.
Fecha de publicaciónfeb-1994
EditorAmerican Society of Plant Biologists
CitaciónPLANT PHYSIOLOGY , Vol 104, Issue 2 453-459
ResumenA cDNA encoding soybean (Glycine max [L.] Merr) ferric leghemoglobin reductase (FLbR), an enzyme that is postulated to play and important role in maintaining leghemoglobin in its functional ferrous state, has been cloned and characterized. A group of highly degenerate oligonucleotides deduced from the N-terminal amino acid sequence of FLbR was used to prime the polymerase chain reaction (PCR) on soybean nodule mRNA and cDNA. A full-length clone of FLbR cDNA was isolated by screening a [lambda]gt11 soybean nodule cDNA library using the specific PCR-amplified FLbR cDNA fragment as a probe. The cDNA contained about 1.8 kb and had a coding sequence for 523 amino acids with a predicted molecular mass of 55,729 D, which included a putative 30-residue signal peptide and a 493-residue mature protein. Computer-aided analysis of the deduced FLbR amino acid sequence showed considerable homology (varied from 20-50% with enzymes and species) to dihydrolipoamide dehydrogenase (EC, glutathione reductase (EC, mercuric reductase (EC, and trypanothione reductase (EC in a superfamily of pyridine nucleotide-disulfide oxidoreductases from various organisms. Northern blot analysis using FLbR cDNA as a probe showed that the FLbR gene was expressed in soybean nodules, leaves, roots, and stems, with a greater level of expression in nodules and leaves than in roots and stems. Southern blot analysis of the genomic DNA showed the presence of two homologous FLbR genes in the soybean genome.
DescripciónPublished as Joumal Series No. 10405, Agricultura1 Research Division, University of Nebraska.
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